Stevia extract containing selected steviol glycosides as flavor, salty and sweetness profile modifier

ABSTRACT

Stevia  extracts with selected major steviol glycosides (Reb A, stevioside, Reb D, Reb C) and minor steviol glycosides and glycosylated diterpene derivative plant molecules, derived from  Stevia rebaudiana  plant are found to improve the perception of flavor and taste perception, which includes the sweet, savory and salty perception in a wide range of food and beverage applications.

RELATED APPLICATIONS

This application is a divisional application of U.S. patent applicationSer. No. 15/619,107, filed on Jun. 9, 2017, which is acontinuation-in-part application of U.S. patent application Ser. No.14/896,022, filed on Dec. 4, 2015, which is a national phase applicationof International Application No. PCT/US2014/041548, filed on Jun. 9,2014, and claims the benefit of priority to U.S. Patent Application No.61/832,451, filed on Jun. 7, 2013, and U.S. Patent Application No.61/942,331, filed on Feb. 20, 2014, the contents of which applicationsare incorporated by reference herein in their entirety.

FIELD OF THE INVENTION

The invention relates to the use of Stevia extracts as flavor modifiersthat contain mixtures of steviol glycosides extracted from Steviarebaudiana plant. This invention also relates to the application of theabove-said Stevia extracts as sweetness profile modifier, not asweetener, with other natural and artificial sweeteners. This inventionalso relates to the production and use of the above-mentioned Steviaextracts that can be used as flavor and sweetness profile modifier whenused in food, beverage, and pharmaceutical products.

BACKGROUND

High intensity sweeteners possess sweetness level many times exceedingthat of sucrose. They are essentially non-caloric and used widely inmanufacturing of diet and reduced calorie food. Although natural caloricsweetener such as sucrose, fructose, and glucose provide the mostdesirable taste to consumers, they are caloric. High intensitysweeteners do not affect the blood glucose level and provide little orno nutritive value.

However, high intensity sweeteners that generally are used assubstitutes for sucrose possess taste characteristics different thanthat of sugar, such as sweet taste with different temporal profile,maximal response, flavor profile, mouthfeel, and/or adaptation behaviorthan that of sugar. For example, the sweet taste of some high-potencysweeteners is slower in onset and longer in duration than that of sugarand thus changes the taste balance of a food composition. Because ofthese differences, usage of high-potency sweetener in replacing such abulk sweetener as sugar in a food or beverage causes imbalance intemporal and/or flavor profile. If the taste profile of high-potencysweeteners could be modified to impart desired taste characteristics, itcan provide low calorie beverages and food products with tastecharacteristics more desirable for consumers. To attain the sugar-liketemporal and/or flavor profile, several ingredients have been suggestedin different publications.

Non-limiting examples of synthetic sweeteners include sucralose,potassium acesulfame, aspartame, alitame, saccharin, neohesperidindihydrochalcone synthetic derivatives, cyclamate, neotame, dulcin,suosan,N—[N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-L-α-aspartyl]-L-phenylalanine1-methyl ester,N—[N-[3-(3-hydroxy-4-methoxyphenyl)-3-methylbutyl]-L-α-aspartyl]-phenylalanine1-methyl ester,N—[N-[3-(3-methoxy-4-hydroxyphenyl)propyl]-L-α-aspartyl]-phenylalanine1-methyl ester, salts thereof, and the like.

Non-limiting examples of natural high intensity sweeteners includeStevioside, Rebaudioside A, Rebaudioside B, Rebaudioside C, RebaudiosideE, Rebaudioside F, Steviolbioside, Dulcoside A, Rubusoside, mogrosides,brazzein, neohesperidin dihydrochalcone (NHDC), glycyrrhizic acid andits salts, thaumatin, perillartine, pernandulcin, mukuroziosides,baiyunoside, phlomisoside-I, dimethyl-hexahydrofluorene-dicarboxylicacid, abrusosides, periandrin, carnosiflosides, cyclocarioside,pterocaryosides, polypodoside A, brazilin, hernandulcin, phillodulcin,glycyphyllin, phlorizin, trilobatin, dihydroflavonol,dihydroquercetin-3-acetate, neoastilibin, trans-cinnamaldehyde, monatinand its salts, selligueain A, hematoxylin, monellin, osladin,pterocaryoside A, pterocaryoside B, mabinlin, pentadin, miraculin,curculin, neoculin, chlorogenic acid, cynarin, siamenoside and others.

High intensity sweeteners can be derived from the modification ofnatural high intensity sweeteners, for example, by fermentation,enzymatic treatment, or derivatization.

A growing number of consumers perceive the ability to control theirhealth by enhancing their current health and/or hedging against futurediseases. This creates a demand for food products with enhancedcharacteristics and associated health benefits, specifically a food andconsumer market trend towards “whole health solutions” lifestyle. Theterm “natural” is highly emotive in the world of sweeteners and has beenidentified as one of key trust, along with “whole grains”,“heart-healthy” and “low-sodium”. ‘Natural’ term is closely related to‘healthier’.

Stevia rebaudiana is a perennial shrub of the Asteraceae (Compositae)family native to certain regions of South America. The leaves of theplant contain from 10 to 20% of diterpene glycosides, which are around150 to 450 times sweeter than sugar. The leaves have been traditionallyused for hundreds of years in Paraguay and Brazil to sweeten localbeverages, foods and medicines.

At present there are more than 230 Stevia species with significantsweetening properties. The plant has been successfully grown under awide range of conditions from its native subtropics to the cold northernlatitudes.

Steviol glycosides have zero calories and can be used wherever sugar isused. They are ideal for diabetic and low calorie diets. In addition,the sweet steviol glycosides possess functional and sensory propertiessuperior to those of many high potency sweeteners.

The extract of Stevia rebaudiana plant contains a mixture of differentsweet diterpene glycosides, which have a single base—steviol and differby the presence of carbohydrate residues at positions C13 and C19. Theseglycosides accumulate in Stevia leaves and compose approximately 10%-20%of the total dry weight. Typically, on a dry weight basis, the fourmajor glycosides found in the leaves of Stevia are Dulcoside A (0.3%),Rebaudioside C (0.6%), Rebaudioside A (3.8%) and Stevioside (9.1%).Other glycosides identified in Stevia extract include Rebaudioside B, C,D, E, and F, Steviolbioside and Rubusoside (FIG. 1).

The chemical structures of the diterpene glycosides of Stevia rebaudianaare presented in FIG. 1. The physical and sensory properties are wellstudied only for Stevioside and Rebaudioside A. The sweetness potency ofStevioside is around 210 times higher than sucrose, Rebaudioside Aaround 300 times, and Rebaudioside C and Dulcoside A around 30 times.The Stevia extract containing Rebaudioside A and Stevioside as majorcomponents showed sweetness potency around 250 times. Rebaudioside A andRebaudioside D are considered to have most favorable sensory attributesof all major Steviol Glycosides (TABLE 1).

TABLE 1 Mol. Solubility in Relative Name Formula T_(Melt), ° C. Weightwater, % sweetness Quality of taste Steviol C₂₀H₃₀O₃ 212-213 318.45 NDND Very bitter Steviolmonoside C₂₆H₄₀O₈ ND 480.58 ND ND ND SteviosideC₃₈H₆₀O₁₈ 196-198 804.88 0.13 210 Bitter Rebaudioside A C₄₄H₇₀O₂₃242-244 967.01 0.80 200-400 Less Bitter Rebaudioside B C₃₈H₆₀O₁₈ 193-195804.88 0.10 150 Bitter Rebaudioside C C₄₄H₇₀O₂₂ 215-217 951.01 0.21 30Bitter Rebaudioside D C₅₀H₈₀O₂₈ 248-249 1129.15 1.00 220 Like sucroseRebaudioside E C₄₄H₇₀O₂₃ 205-207 967.01 1.70 170 Like sucroseRebaudioside F C₄₃H₆₈O₂₂ ND 936.99 ND ND ND Dulcoside A C₃₈H₆₀O₁₇193-195 788.87 0.58 30 Very bitter Steviolbioside C₃₂H₅₀O₁₃ 188-192642.73 0.03 90 Unpleasant Rubusoside C₃₂H₅₀O₁₃ ND 642.73 ND 110 Verybitter

In addition to the commercially known steviol glycosides (Table 1),several new steviol glycosides (glycosylated diterpene) have been foundin Stevia leaf extracts, as shown in Table 2a.

TABLE 2a Summary of formula and R-groups of identified steviolglycosides (see FIG. 1 for backbone structure) Trivial Mol. # Commonname formula Wt. R₁ R₂ Reference 1. Steviol + Glucose (SvGn) 1.1Steviolmonoside SvG1   481 H Glcβ1- Ohta et al. (2010) 1.2Steviolmonoside SvG1   481 Glcβ1- H Gardena et A al. (2010) 1.3Rubusoside SvG2   643 Glcβ1- Glcβ1- Ohta et al. (2010) 1.4Steviolbioside SvG2   643 H Glcβ(1-2)Glcβ1- Kohda et al. (1976) 1.5Stevioside SvG3   805 Glcβ1- Glcβ(1-2)Glcβ1- Bridel & Lavielle (1931)1.6 Stevioside A SvG3   805 Glcβ(1- Glcβ1- Wu et al. 2)Glcβ1- (2012) 1.7Rebaudioside B SvG3   805 H Glcβ(1- Kohda et 2)[Glcβ(1- al. (1976)3)]Glcβ1- 1.8 Rebaudioside G SvG3   805 Glcβ1- Glcβ(1-3)Glcβ1- Ohta etal. (2010) 1.9 Stevioside B SvG3   805 Glcβ(1- Glcβ1- Chaturvedula3)Glcβ1- & Zamora (2014) 1.10 Rebaudioside E SvG4   967 Glcβ(1-Glcβ(1-2)Glcβ1- Sakamoto 2)Glcβ1- et al. (1977a) 1.11 Rebaudioside ASvG4   967 Glcβ1- Glcβ(1- Kohda et 2)[Glcβ(1- al. (1976) 3)]Glcβ1- 1.12Rebaudioside A2 SvG4   967 Glcβ1- Glcβ(1- Chaturvedula 6)Glcβ(1- &Prakash 2)Glcβ1- (2011d) 1.13 Rebaudioside D SvG5 1 129 Glcβ(1- Glcβ(1-Sakamoto 2)Glcβ1- 2)[Glcβ(1- et al. 3)]Glcβ1- (1977a) 1.14 RebaudiosideI SvG5 1 129 Glcβ(1- Glcβ(1- Ohta et al. 3)Glcβ1- 2)[Glcβ(1- (2010)3)]Glcβ1- 1.15 Rebaudioside L SvG5 1 129 Glcβ1- Glcβ(1- Ohta et al.6)Glcβ(1- (2010) 2)[Glcβ(1- 3)]Glcβ1- 1.16 Rebaudioside Q2 SvG5 1 129Glcα(1- Glcβ(1-2)Glcβ1- Chaturvedula 2)Glcα(1- & Prakash 4)Glcβ1-(2011c) 1.17 Rebaudioside Q SvG5 1 129 Glcβ1- Glcα(1- — 4)Glcβ(1-2)[Glcβ(1- 3)]Glcβ1- 1.18 Rebaudioside I2 SvG5 1 129 Glcβ1- Glcα(1-Chaturvedula 3)Glcβ(1- et al. 2)[Glcβ(1- (2011c) 3)]Glcβ1- 1.19Rebaudioside Q3 SvG5 1 129 Glcβ1- Glcα(1- Chaturvedula 4)Glcβ(1- et al.3)[Glcβ(1- (2011c) 2)]Glcβ1- 1.20 Rebaudioside I3 SvG5 1 129 Glcβ(1-Glcβ(1-2)Glcβ1- Chaturvedula 2)[Glcβ(1- et al. 6)]Glcβ1- (2011c) 1.21Rebaudioside M SvG6 1 291 Glcβ(1- Glcβ(1- Ohta et al. 2)[Glcβ (1-2)[Glcβ(1- (2010) 3)]Glcβ1- 3)]Glcβ1- 2. Steviol + Rhamnose + Glucose(SvR1Gn) 2.1 Dulcoside A SvR1G2   789 Glcβ1- Rhaα(1-2)Glcβ1- Kobayashiet al. (1977) 2.2 Dulcoside B SvR1G2   789 H Rhaα(1- Ohta et al.2)[Glcβ(1- (2010) 3)]Glcβ1- 2.3 Rebaudioside C SvR1G3   951 Glcβ1-Rhaα(1- Sakamoto 2)[Glcβ(1- et al. 3)]Glcβ1- (1977b) 2.4 RebaudiosideSvR1G3   951 Rhaα(1- Glcβ(1-2)Glcβ1- Purkayastha C2^(a) 2)Glcβ1- (2016)2.5 Rebaudioside S SvR1G3   951 Rhaα(1- Glcα (1- Ibrahim et 2)Glcβ1-2)Glcβ1- al (2016) 2.6 Rebaudioside H SvR1G4 1 112 Glcβ1- Glcβ(1- Ohtaet al. 3)Rhaα(1- (2010) 2)[Glcβ(1- 3)]Glcβ1- 2.7 Rebaudioside K SvR1G4 1112 Glcβ(1- Rhaα(1- Ohta et al. 2)Glcβ1- 2)[Glcβ(1- (2010) 3)]Glcβ1- 2.8Rebaudioside J SvR1G4 1 112 Rhaα(1- Glcβ(1- Ohta et al. 2)Glcβ1-2)[Glcβ(1- (2010) 3)]Glcβ1- 2.9 Rebaudioside N SvR1G5 1 274 Rhaα(1-Glcβ(1- Ohta et al. 2)[Glcβ(1- 2)[Glcβ(1- (2010) 3)]Glcβ1- 3)]Glcβ1-2.10 Rebaudioside O SvR1G6 1 436 Glcβ(1- Glcβ(1- Ohta et al. 3)Rhaα(1-2)[Glcβ(1- (2010) 2)[Glcβ(1- 3)]Glcβ1- 3)]Glcβ1- 2.11 RebaudiosideSvR1G6 1 436 Glcβ(1- Glcβ(1- Purkayastha O2^(a) 4*)Rhaα(1- 2)[Glcβ(1-(2016) 2)[Glcβ(1- 3)]Glcβ1- 3)]Glcβ1- 2.12 Rebaudioside SvR1G4 1 112Glcβ(1- Rhaα(1- Purkayastha K2^(a) 6)Glcβ1- 2)[Glcβ(1- (2016) 3)]Glcβ1-3. Steviol + Xylose + Glucose (SvX1Gn) 3.1 Stevioside F SvX1G2   775Glcβ1- Xylβ(1-2)Glcβ1- Chaturvedula & Prakash (2011b) 3.2 Rebaudioside FSvX1G3   937 Glcβ1- Xylβ(1- Starratt et 2)[Glcβ(1- al. (2002) 3)]Glcβ1-3.3 Rebaudioside F2 SvX1G3   937 Glcβ1- Glcβ(1- Chaturvedula 2)[Xylβ(1-& Prakash 3)]Glcβ1- (2011b) 3.4 Rebaudioside F3 SvX1G3   937 Xylβ(1-Glcβ(1-2)Glcβ1- Chaturvedula 6)Glcβ1- et al. (2011d) 3.5 Rebaudioside RSvX1G3   937 Glcβ1- Glcβ(1- Ibrahim et 2)[Glcβ(1-3)] al (2016) Xylβ1-3.6 Rebaudioside SvX1G4 1 099 Xylβ(1- Glcβ(1- Purkayastha U^(a) 2)Glcβ1-2)[Glcβ(1- (2016) 3)]Glcβ1- 3.7 Rebaudioside SvX1G4 1 099 Xylβ(1-Glcβ(1-2)Glcβ1- Purkayastha U2^(a) 2*)[Glcβ(1- (2016) 3)]Glcβ1- 3.8Rebaudioside SvX1G5 1 261 Glcβ(1- Xylβ(1- Purkayastha V^(a) 2)[Glcβ(1-2*)[Glcβ(1- (2016) 3)]Glcβ1- 3)]Glcβ1- 3.9 Rebaudioside SvX1G5 1 261Xylβ (1- Glcβ(1- Prakash & V2^(a) 2)[Glcβ(1- 2)[Glcβ(1- Chaturvedula3)]Glcβ1- 3)]Glcβ1- (2013) 4. Steviol + Arabinose + Glucose (SvA1Gn) 4.1Rebaudioside SvA1G4 1 098 Glcβ(1- Glcβ(1-2)Glcβ1- Purkayastha W^(a)2)[Araβ(1- (2016) 3*)]Glcβ1 4.2 Rebaudioside SvA1G4 1 098 Araβ(1-Glcβ(1- Purkayastha W2^(a) 2*)Glcβ1 2)[Glcβ(1- (2016) 3)]Glcβ1- 4.3Rebaudioside SvA1G4 1 098 Araβ(1- Glcβ(1- Purkayastha W3^(a) 6)Glcβ1-2)[Glcβ(1- (2016) 3)]Glcβ1- 4.4 Rebaudioside SvA1G5 1 260 Glcβ(1-Glcβ(1- Purkayastha Y^(a) 2)[Araβ(1- 2)[Glcβ(1- (2016) 3*)]Glcβ13)]Glcβ1- 5. Steviol + Fructose + Glucose (SvF1Gn) 5.1 Rebaudioside A3SvF1G3   967 Glcβ1- Glcβ(1- Chaturvedula 2)[Fruβ(1- et al. 3)]Glcβ1-(2011b) 6. Steviol + galactose + Glucose (SvGa1Gn) 6.1 RebaudiosideSvGa1 1 128 Galβ(1- Glcβ(1- Purkayastha T^(a) G4 2*)Glcβ1 2)[Glcβ(1-(2016){circumflex over ( )} 3)]Glcβ1- 7. Steviol + de-oxy glucose +Glucose (SvdG1Gn) 7.1 Stevioside D SvdG1G2   789 Glcβ1- 6-deoxyGlcβ(1-Chaturvedula 2)Glcβ1- & Prakash (2011a) 7.2 Stevisoide E SvdG1G3   951Glcβ1- 6-deoxyGlcβ(1- Chaturvedula 2)[Glcβ(1- & Prakash 3)]Glcβ1-(2011a) 7.3 Stevioside E2 SvdG1G3   951 6- Glcβ(1- ChaturveduladeoxyGlcβ1- 2)[Glcβ(1- et al. 3)]Glcβ1- (2011e)

Besides diterpene glycosides, a number of flavonoids, labdane diterpene,triterpenes, sterols, and volatile oils have also been found in theextracts of Stevia rebaudiana, collectively referred to as plantmolecules, as shown in Table 2b.

TABLE 2b Chemical Classes Chemical Components Monoterpenoids BorneolDiterpenoids Austroinulin, 6-0-acetyl austroinulin, 6-acetylaustroinulin 7-0-acetyl austroinulin, Sterebin A, B, C, D, E, F, G, H,Jhanol Triterpenoids Amyrin beta acetate Sesquiterpenes α-bergamotene,Bisabolene, β-bourbonene, δ-cadinene, γ-cadinene Essential oilsβ-caryophyllene, Trans β-tarnesene, α-humulene, δ-cadiene caryophylleneoxide, Nerolidol, Linalol, α-terpineol, Terpinen-4-ol Sterol derivativesStigmasterol , β-sitosterol, Campesterol FlavonoidsGlucosy1-4′-O-apigenin, Glucosyl-7-O-luteolin, Rhamnosyl-3-O-kaempferol, Quercetin, Glucosyl-3-O-quercetin, Arabinosyl-3-O-quercetin, 5,7,3′-methoxyflavone, 3,6,4′-methoxyflavone, Centaureidin,avicularin

All steviol glycosides provide sweetness and other taste attributes at ahigher than certain threshold level of concentrations in water. Belowthe threshold level of concentration, the steviol glycoside componentsand their mixtures as found in a typical non-limiting Stevia extract asshown below has no recognizable sweetness taste. But such Stevia extractbelow the threshold level of significant sweetness recognition showremarkable characteristics of sweet and flavor profile modification infood and beverage applications.

This invention relates to use of the following Stevia extracts (Table 3)with the varying level of different steviol glycosides and other Steviaplant-derived glycosides, the combination of which contributes nosignificant sweetness but modifies flavor and sweetness profile atcertain concentration in typical food and beverage applications.

TABLE 3 Minor Steviol Glycosides Steviol Glycosides*, % and relatedStevia Reb Dulco- Rubu- Steviolbio- TSG* plant Extracts Reb A SteviosideReb D F Reb C side A soside Reb B side Reb E Reb N Reb O (%) moleculesPCS-5001 10-20 4-12 1-4 1-5 10-25 1-5 1-4 0.5-5 0.5-5 1-4 0.5-4 0.5-445-65 35-50 PCS-1015 18-25 5-10 8-20 0-1  1-3 0-1 0-1 0.5-5   0-1 2-6  4-8   3-8 55-65 35-45 PSB-5005 15-30 3-12 1-10 0-5  5-15 0-5 0-5   0-8  0-5 0-5   0-6   0-5 45-70 30-55% *TSG or Total Steviol Glycosidescontain the Steviol Glycosides that are recognized by Codex Alimentarius(a commission of FAO and WHO) and major regulatory authorities

The present invention also relates to the Stevia extracts that containmajor steviol glycosides (Table 3) and other minor steviol glycosidesand glycosylated diterpene derivatives (water soluble molecules). Thenon-limiting examples of such minor molecules are Reb E, Reb G, Reb H,Reb I, Reb K, Reb L, Reb M, Reb N, Reb O (M. Ohta, S. Sasa, A. Inoue, etal. “Characterization of Novel Steviol Glycosides from Leaves of Steviarebaudiana Morita.” J. Appl. Glycosci., 57, 1 99-209 (201 0)).

The present invention is also directed to a method of making a specificStevia extract composition, including: extracting steviol glycosides andother water soluble molecules from leaves of a Stevia rebaudiana plant,and separating the excess steviol glycosides than the amount and type ofsteviol glycosides required to contribute the taste and flavor modifyingcharacteristics of the Stevia extract.

This invention combine the different natural sweeteners, especiallysteviol glycosides in certain proportion along with other water solublemolecules to provide enhanced sweetness and flavor profile in food andbeverage application, which can be blended with other natural caloricsweeteners to impart more desirable sweetness profile. Non-limitingexamples of caloric sweeteners include dextrose, fructose, sucrose,maltose, lactose, corn syrup, gluco-syrup derived from differentcarbohydrates, cane syrup, flavored sugar, honey, molasses,

This invention combine the different natural sweeteners, especiallysteviol glycosides in certain proportion along with other water solublemolecules to provide enhanced sweetness and flavor profile in food andbeverage application, which can be blended with other naturalnon-caloric sweeteners to impart more desirable sweetness profile.Non-limiting examples of natural high intensity sweeteners includesteviol glycosides, brazzein, monatin and its salt, neohesperidindihydrochalcone (NHDC), glycyrrhizic acid and its salts, thaumatin,mogrosides and lu han guo extracts, perillartine, mabinlin, pentadin,miraculin, curculin, neoculin, chlorogenic acid, cynarin, siamenosideand others.

This invention combine the different natural sweeteners, especiallysteviol glycosides in certain proportion along with other water solublemolecules to provide enhanced sweetness and flavor profile in food andbeverage application, which can be blended with other syntheticnon-caloric sweeteners to impart more desirable sweetness profile.Non-limiting examples of synthetic sweeteners include sucralose,potassium acesulfame, aspartame, alitame, advantame, saccharin,neohesperidin dihydrochalcone synthetic derivatives, cyclamate, neotame,dulcin, suosan,N—[N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-L-α-aspartyl]-phenylalanine1-methyl ester,N—[N-[3-(3-hydroxy-4-methoxyphenyl)-3-methylbutyl]-α-aspartyl]-L-phenylalanine1-methyl ester,N—[N-[3-(3-methoxy-4-hydroxyphenyl)propyl]-L-α-aspartyl]-L-phenylalanine1-methyl ester, salts thereof, and the like.

BRIEF SUMMARY OF THE INVENTION

The present invention is directed to a taste and flavor modifyingcomposition. The composition includes different steviol glycosides withother water soluble molecules derived from Stevia leaf, such asnon-limiting examples of plant glycosides, flavonoids, labdanediterpene, triterpenes, which can modify the intensity of a taste and/ora flavor in a food or beverage product.

The present invention is also directed to a food or beverage producthaving an intense taste and flavor profile, wherein the food or beverageproduct includes a taste and flavor modifying composition comprising theStevia extract of steviol glycosides and water soluble molecules derivedfrom Stevia plant. A wide range of food and beverage products, such as,but not limited to, carbonated soft drinks, fruit juices, dairy foods,dairy beverages, baked goods, cereal products, snack foods, and tabletop sweeteners, may be made in accordance with the present invention.The taste and flavor profile of a food or beverage product including ataste and flavor modifying composition, wherein the taste and flavormodifying composition comprising the Stevia extract of steviolglycosides and water soluble molecules derived from Stevia plant, may bemore intense than a comparative taste and flavor profile of acomparative food or beverage product which does not include the tasteand flavor modifying composition. Moreover, the mouthfeel and overalltaste perception of a food or beverage product including the taste andflavor modifying composition, wherein the taste and flavor enhancingcomposition includes the complex mixture of steviol glycosides and watersoluble molecules, may be improved in relation to a mouthfeel andoverall taste perception of a comparative food or beverage product whichdoes not include the taste and flavor enhancing composition.

The present invention is further directed to a method of increasing thetaste and flavor intensity of a food or beverage product, including thestep of adding a taste and flavor enhancing composition to the food orbeverage product, wherein the taste and flavor modifying compositioncomprising the Stevia extract of steviol glycosides and water solublemolecules derived from Stevia plant. The present invention is alsodirected to a method of improving the organoleptic properties of a foodor beverage product including a high fructose syrup, including the stepof adding the taste and flavor modifying composition to the food orbeverage product. For example, adding the taste and flavor modifyingcomposition may cause the high fructose syrup, such as high fructosecorn syrup, to taste more like sugar. Also, if the high fructose syrupis high fructose corn syrup 42 (HFCS 42), adding the taste and flavorenhancing composition may cause the HFCS 42 to taste more like highfructose corn syrup 55 (HFCS 55).

The present invention is further directed to a method of increasing thetaste and flavor intensity of a medical food and pharma product,including the step of adding a taste and flavor modifying composition tothe food or beverage product, wherein the taste and flavor modifyingcomposition comprising the Stevia extract of selected steviol glycosidesand water soluble molecules derived from Stevia plant. The presentinvention is also directed to a method of improving the organolepticproperties of a medical food or pharma product containing functionalfood ingredients like vitamins, minerals and amino acids, including thestep of adding the taste and flavor modifying composition to the food orbeverage product. For example, adding the taste and flavor modifyingcomposition may cause the off-taste due to vitamins, mineral, aminoacids and other non-limiting functional ingredients, to improve tasteand palatability.

The present invention is also directed to a method of making a taste andflavor enhancing composition, including: extracting steviol glycosidesand other water soluble molecules from leaves of a Stevia rebaudianaplant, and separating the excess steviol glycosides than the amount andtype of steviol glycosides required to contribute the taste and flavormodifying characteristics of the Stevia extract.

The foregoing has outlined rather broadly the features and technicaladvantages of the present invention in order that the detaileddescription of the invention that follows may be better understood.Additional features of the invention which form the subject of theclaims of the invention will be described hereinafter. It should beappreciated by those skilled in the art that the specific embodimentsdisclosed may be readily utilized as a basis for modifying or designingother methods or structures for carrying out the same purposes of thepresent invention. It should also be realized by those skilled in theart that such equivalent constructions do not depart from the spirit andscope of the invention as set forth in the appended claims. The novelfeatures which are believed to be characteristic of the invention, bothas to its organization and method of operation, together with furtherobjects and advantages will be better understood from the followingdescription.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the chemical structure of the diterpene glycosides ofStevia rebaudiana.

FIG. 2 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to a cola flavoredcarbonated soft drink.

FIG. 3 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to an iced teabeverage.

FIG. 4 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to an iced teabeverage.

FIG. 5 is a bar graph showing the effect of Stevia extract on the flavorprofile of roasted peanuts.

FIG. 6 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to tomato ketchup.

FIG. 7 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to chocolate milk.

FIG. 8 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to poppy seed muffins.

FIG. 9 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to tortilla chips.

FIG. 10 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to beef jerky.

FIG. 11 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to brown gravy.

FIG. 12 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to chocolate milk.

FIG. 13 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to vanilla custard.

FIG. 14 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to chocolate milk.

FIG. 15 is a bar graph showing the modification of flavor and sweetnessprofiles caused by the addition of Stevia extract to vanilla yogurt.

DETAILED DESCRIPTION

Embodiments of the present invention are described in the followingexamples.

EXAMPLES Example 1A: Detection of Concentration Threshold for SweetnessRecognition

To detect the sweetness recognition level of PCS-5001, PCS 1015 and PSB5005 (Stevia extract), the test method outlined by Harman, et al (FoodTechnology, November 2013) was used with ten trained panelists that havebeen previously qualified for their taste acuity and trained in the useof a sweetness intensity rating scale. The panelists evaluated a seriesof aqueous solutions of sucrose and the Stevia extract (PCS-5001,PCS-1015, or PSB 5005) at room temperature; the sucrose solutions of1.5% concentration and the Stevia extract solutions with concentrationsranging between 100 and 120 ppm for PCS-5001,70-80 ppm for PCS-1015, and60-70 ppm for PSB 5005 were prepared with filtered water. The objectiveof the test was to determine the sweetness recognition level of theStevia extract. The evaluations were done in triplicate using the samepanelists so that a total of 30 values were generated for each averagedata point.

The samples were coded and presented in random order to panel members totaste and determine which sample was sweeter (ASTM E2164-08: StandardMethod for Directional Difference Test). Panelists were asked to focusonly on sweet attribute of those samples and to use warm water and saltsolution in order to cleanse the palate between samples.

The results were tallied and significance was calculated by SIM 2000(Sensory Computer System, NJ). Results are presented in Table 4. Theoverall sweetness of those samples was barely detectable. The 2-AFCshows that 100 ppm PCS-5001, 70 ppm of PCS-1015 and 60 ppm of PSB 5005solutions were the least sweet samples and were significantly less sweetthen the 1.5% sugar control. The sample with 120 ppm PCS-5001 and 80 ppmPCS-1015 were the sweetest samples showing significantly highersweetness than the 1.5% sugar control (Table 4). The recognitionthreshold concentration of STEVIA EXTRACT (PCS-5001) in water wasdetermined to be 100 ppm. The recognition threshold concentration ofSTEVIA EXTRACT (PCS-1015) in water was determined to be 70 ppm. Thesweetness recognition threshold of STEVIA EXTRACT (PSB 5005) in waterwas determined to be 60 ppm.

TABLE 4 Sweetness perception of Stevia Extract in differentconcentration against 1.5% sugar solution. Comparison of sweetnessperception of STEVIA Sugar solution Stevia Extract EXTRACT in water(1.5%) sweeter? solution sweeter? P-Value Significance PCS-5001: 100ppm, N=30 23 7 0.0052 *** PCS-5001: 110 ppm, N=30 20 10 0.0987 ** PCS-5001: 120 ppm, N=30 9 21 0.0457 *** PCS-1015:  70 ppm, N=30 26 40.0001 *** PCS-1015:  80 ppm, N=30 5 25 0.0003 *** PSB-5005:  60 ppm,N=30 24 6 0.0014 *** PSB-5005:  70 ppm, N=30 19 11 0.2005 NS 

Example 1B: Sweetness Detection of Concentration Threshold for SweetnessDetection

The ten panel members evaluated a series of lemon-lime flavoredcarbonated soft drink (CSD) sweetened with sucrose and STEVIA EXTRACT atroom temperature; the evaluations were done in triplicate using the samepanelists so that at least 30 values were generated for each averagedata point. The lemon lime flavored carbonated soft drink control samplehad 1.5% sucrose concentration and the test sample contained STEVIAEXTRACT (PCS-5001) with concentrations at 110 and 120 ppm or STEVIAEXTRACT (PCS-1015) with concentrations of 70 and 90 ppm. Otheringredients in the CSD samples were citric acid, lemon-lime flavor,sodium benzoate, potassium citrate and xanthan gum. The objective of thetest was to determine the sweetness detection limit of STEVIA EXTRACT.Tests were conducted as outlined in Example 1A.

The samples with 120 ppm PCS-5001(STEVIA EXTRACT) and 90 ppmPCS-1015(STEVIA EXTRACT) showed no significant difference in sweetnessthan the 1.5% sugar control. The recognition threshold concentration ofPCS-5001(STEVIA EXTRACT) in a lemon-lime flavored carbonated soft drinkwater was determined to be 110 ppm. The recognition thresholdconcentration of PCS-1015 (STEVIA EXTRACT) in a lemon-lime flavoredcarbonated soft drink water was determined to be 70 ppm. Results areshown in table 5.

TABLE 5 Sweetness perception of STEVIA EXTRACT in differentconcentrations against 1.5% sugar solution in a typical carbonated softdrink (CSD) Sweetness perception of CSD sample with CSD sample withSIEVIA EXTACT in CSD Sugar sweeter? Stevia Sweeter? P-Value SignificancePCS-5001: 110 ppm, N=30 23 7 0.0052 *** PCS-5001: 120 ppm, N=36 20 160.677 NS PCS-1015:  70 ppm, N=30 21 9 0.0428 *** PCS-1015:  90 ppm, N=3012 18 0.3616 NS

Example 2: Effect of Stevia Extract on Flavor Modification in a TypicalCarbonated Soft Drink Application

A cola flavored carbonated soft drink was developed to evaluate theeffect of PCS-5001 and PCS-1015 (Stevia extract) on the sweetness andflavor profile of the beverage that was sweetened with sugar and Steviasweetener to achieve 30% sugar reduction (Table 6). The samples with andwithout PCS-5001 or PCS-1015 were evaluated by thirty consumer panelmembers, who assigned relative values to each sample for overall Liking,sweetness, vanilla flavor, brown note, and aftertaste on a 10-ptcontinuous intensity scale as outlined in Table 7.

TABLE 6 Cola flavored Soft drink for sensory evaluation Control: Test:30% Sugar Test: 30% Sugar COLA BEVERAGE 30% Reduction with Reductionwith FORMULA Sugar Reduction PCS-5001 PCS-1015 Water 91.68 91.67 91.67Sugar 7.89 7.89 7.89 Cola Flavor-Flavor Systems 0.375 0.375 0.375Phosphoric Acid 85% 0.0333 0.0333 0.0333 Caffeine 0.0100 0.0100 0.0100Steviol glycoside 0.0100 0.0100 0.0100 PCS-5001 0.0110 PCS-1015 0.0080Total 100 100 100

TABLE 7 Sensory evaluation of Cola flavored carbonated soft drink Natureof Participants: Company employees Number of Sessions 1 Number of 30Participants: Test Design: Balanced, randomized within pair. BlindSensory Test Intensity and acceptance ratings Method: EnvironmentalStandard booth lighting Condition Attributes and Scales: OverallAcceptance on a 10-pt hedonic scale where 10 = Extremely Like and 0 =Extremely Dislike Overall Liking, Sweetness, Vanilla flavor, Brown note,and Sweet Aftertaste. 10-pt continuous intensity scale where 0 =Imperceptible and 10 = Extremely Pronounced Statistical Analysis: ANOVA(by Block) with Post Hoc Duncan's Test Sample Size ~1.5 oz in a clearcapped plastic cup Serving Temperature Refrigerated temperature (~45°F.) Serving/Panelists Samples served simultaneously. Panelistsinstructed to Instruction: read ingredient statement, evaluate eachsample.

FIG. 2 shows the modification of flavor and sweetness profiles caused bythe addition of Stevia extract (PCS-5001). The results indicated thesample containing Stevia extract PCS-5001 and the sample containingPCS-1015 had significantly higher cola flavor, vanilla flavor, brownspice notes and overall liking compared to the control samples (at 95%confidence). The sample containing PCS-5001 had directionally lowerbitterness, and bitter aftertaste intensity compared to the controlsamples (at 90% and 95% confidence respectively). The sample containingPCS-1015 had directionally lower bitterness, and sweet aftertasteintensity compared to the control samples (at 80% confidence). Inaddition, the sample with Stevia extract (PCS-1015) had significantlylower bitter aftertaste compared to the control sample (at 95%confidence).

Example 3: Peach Flavored Tea Beverage for Sensory Evaluation

A peach flavored black tea drink was developed to evaluate the effect ofSTEVIA EXTRACT on the sweetness and flavor profile of the beverage thatwas sweetened with sugar and Stevia sweetener to achieve 30% sugarreduction (Table 8). The samples with and without STEVIA EXTRACT wereevaluated as outlined in EXAMPLE 2 by thirty consumer panel members, whoassigned relative values to sweetness, bitterness, peach flavor, teaflavor, acid intensity, astringency, and aftertaste on 10-pt continuousintensity scale where 0=Imperceptible and 10=extremely pronounced.

TABLE 8 Peach Flavored Tea Beverage samples for sensory evaluationReduce Sugar Reduce Sugar Reduced Sugar Tea with PCS- Tea with PCS- Tea5001 1015 Water 95.71 95.70 95.71 Sucrose 3.850 3.850 3.850 Black TeaPowder 0.275 0.275 0.275 Citric Acid 0.0880 0.0880 0.0880 Peach Flavor0.0330 0.0330 0.0330 Sodium Citrate 0.0150 0.0150 0.0150 PotassiumSorbate 0.0150 0.0150 0.0150 Steviol Glycoside 0.0140 0.0140 0.0140Stevia Extract PCS-5001 0.0120 Stevia Extract PCS-1015 0.0080 XanthanGum-TIC 0.0013 0.0013 0.0013

FIG. 3 shows the modification of flavor and sweetness profilescontributed by the addition of STEVIA EXTRACT (PCS-5001) in peachflavored ice tea beverage. The results indicated that the test samplecontaining PCS-5001 had significantly higher peach flavor, and overallliking (at 95%, confidence). The sample containing PCS-5001 hadsignificantly lower astringency than the control sample (at 95%confidence). The results shown in FIG. 4 indicated that the test samplecontaining PCS-1015 had significantly higher peach flavor, black teaflavor, and overall liking (at 95%, confidence). The sample PCS-1015also had significantly lower astringency, sweet intensity, bitterintensity, and bitter aftertaste than the Control sample (at 95%confidence). In addition, the PCS-1015 sample had lower sweet aftertasteintensity than the Control sample at 90% confidence).

Example 4: Effect of Stevia Extract on Flavor Modification of SavoryApplications

A seasoning blend was developed to determine the flavor modificationeffect of Stevia extract in a seasoning blend on reduced sugar roastedpeanut samples. Thirty consumer panel members evaluated two samples ofthe peanuts for overall acceptance and attribute intensities (overallflavor, saltiness, sweetness, smoke flavor, spice/heat intensity, peanutflavor, chili powder flavor, bitterness and lingering sweet aftertasteintensity). The two samples (Table 9) included: 1) 50% sugar reducedcontrol sample containing Stevia glycosides, and 2) 50% reduced sugartest sample containing steviol glycoside and Stevia extract, PCS-5001 orPCS-1015.

The objective of the test was to determine if the addition of Steviaextract affects the flavor profile of a savory snack food. The resultsindicated that the addition of PCS-5001 at 110 ppm and PCS-1015 at 70ppm provided flavor modification (FIG. 5). The test samples containing110 ppm PCS-5001 had significantly higher salt intensity, smoke flavor,and bitter intensity compared to the control (95% confidence). The testsample also had lower sweet intensity than the control (95% confidence).In addition, the test sample containing Stevia extract had directionallyhigher spice and chili notes (90% confidence). The test samplecontaining PCS-1015 had significantly higher salt intensity than thecontrol sample (at 95% confidence). The test sample showed an increasein heat/spice intensity, and chili flavor compared to the control.

TABLE 9 Effect of STEVIA EXTRACT on snack and seasoning applicationsSteviol Steviol Glycoside + Steviol Glycoside + Glycoside Stevia ExtractStevia Extract Unsalted Peanuts 86.8 86.8 86.8 Vegetable oil 2.93 2.932.93 Sugar 5.88 5.88 5.88 Salt 2.93 2.93 2.93 Chilli powder 0.174 0.1740.174 Cumin powder 0.286 0.286 0.286 Garlic powder 0.156 0.156 0.156Cayenne pepper 0.156 0.156 0.156 Smoke liquid 0.729 0.729 0.729 SteviolGlycoside 0.0243 0.0243 0.0243 PCS-5001 0.0110 PCS-1015 0.0070 Total wt.(g) 100 100 100

TABLE 10 Sensory evaluation of snack and seasoning applications Natureof Participants: Company employees Number of Sessions 1 Number ofParticipants: 30 Test Design: Balanced, randomized within pair. BlindSensory Test Method: Intensity and acceptance ratings EnvironmentalCondition Standard booth lighting Attributes and Scales: OverallAcceptance on a 9-pt hedonic scale where 9 = Like Extremely, 5 = NeitherLike Nor Dislike, and 1 = Dislike Extremely Overall Flavor, Saltiness,Sweetness, Smoke Intensity, Heat/spice intensity, peanut flavor, chilipowder and Aftertaste Intensity (sweet and bitter) on a 10-pt continuousintensity scale where 0 = Imperceptible and 10 = Extremely PronouncedOpen Ended General Comments Statistical Analysis: ANOVA (by Block) withPost Hoc Duncan's Test Sample Size ~1.5 oz in a clear capped plastic cupServing Temperature Room temperature (~70° F.) Serving/Panelists Samplesserved simultaneously. Panelists evaluate each Instruction: sample once.

Example 5: Flavor Modification of Sauce and Vegetable Preparation

A tomato ketchup preparation was developed to determine the flavormodification effect of Stevia extract (PCS-1015). A panel of thirtycompany employees evaluated the overall acceptance and attributeintensities (tomato, onion, vinegar, sweet, saltiness, bitterness andaftertaste) of each sample. The sensory evaluation methodology outlinedin Example 4 was adopted for the sauce samples as presented in Table 11.

TABLE 11 Effect of PCS-1015 (stevia extract) on tomato ketchup SteviolGlycoside Steviol Glycoside w/Stevia Extract Tomato Juice (Sieved)52.4863 52.4793 Tomato Puree 24.6236 24.6236 White Distilled Vinegar11.3454 11.3454 Water 1.5845 1.5845 Sucrose 2.6511 2.6511 Tomato Paste5.8311 5.8311 Onion Powder 0.8649 0.8649 Salt 0.5811 0.5811 Steviolglycoside 0.032 0.032 Stevia Extract (PCS 1015) 0.007 Total 100 100

FIG. 6 shows the modification of flavor and sweetness profiles caused bythe addition of Stevia extract (PCS-1015). The results indicate the testsamples containing Stevia extract, PCS-1015, had a significant increasein herbal notes, and savory (onion/garlic) notes at a 95% confidenceinterval. The test sample containing PCS-1015 had directionally lowerbitterness, bitter aftertaste and overall liking at a 90% confidenceinterval compared to the control sample.

Example 6: Effect of PCS-1015 (Stevia Extract) on Flavor Modification ofDairy Applications

A chocolate flavored dairy beverage was developed to determine theflavor modification effect of Stevia extract (PCS-1015) in dairybeverage. The panel evaluated samples of chocolate milk for overallacceptance and attribute intensities (chocolate flavor, dairy notes,sweetness, bitterness and aftertaste). The two samples (Table 12)included: 1) 50% sugar reduced control sample containing Steviaglycosides, and 2) 50% reduced sugar test sample containing Steviaglycoside and 80 ppm of Stevia extract, PCS-1015.

TABLE 12 Effect of PCS-1015 (stevia extract) on flavored dairy beverage50% Total Sugar 50% Total Sugar Reduction Reduction with stevia withsteviol extract and stevia Dairy Formula glycoside glycoside 2% Reducedfat Milk 96.5803 96.5753 Sugar 2.40 2.40 Cocoa Powder 0.80 0.80Palsgaard 150 ChoMilk 0.20 0.20 Steviol Glycosides 0.0197 0.0197PCS-1015 0.080 Total 100 100

TABLE 13 Sensory evaluation of Dairy beverage Nature of Participants:Company employees Number of Sessions 1 Number of Participants: 30 TestDesign: Balanced, randomized within pair. Blind Sensory Test Method:Intensity and acceptance ratings Environmental Standard booth lightingCondition Attributes and Scales: Overall Acceptance on a 10-pt hedonicscale where 10 = Extremely Like and 0 = Extremely Dislike OverallLiking, sweetness, bitterness, dairy notes, chocolate, and Aftertaste.10- pt continuous intensity scale where 0 = Imperceptible and 10 =Extremely Pronounced Statistical Analysis: ANOVA (by Block) with PostHoc Duncan's Test Sample Size ~1.5 oz. in a clear capped plastic cupServing Temperature Refrigerated temperature (~45° F.) Serving/PanelistsSamples served simultaneously. Panelists instructed to read Instruction:ingredient statement, evaluate each sample.

FIG. 7 shows the modification of flavor and sweetness profiles caused bythe addition of Stevia extract (PCS-1015). The results indicate the 50%sugar reduced sample containing steviol glycoside sweetener and Steviaextract, PCS-1015, had significantly higher chocolate flavor.

Example 7: Effect of Stevia Extract (PCS-5001) on Flavor Modification ofBaked Goods Applications

A lemon poppy seed flavored muffin formulation was developed todetermine the flavor modification effect of Stevia extract (PCS-5001) inbaked good applications. To test the contribution of PCS-5001 in bakedgoods, lemon flavored poppy seed muffins were baked with a 45% sugarreduced formulation with steviol glycoside as control, and sugar reducedformulation with steviol glycoside and Stevia extract (PCS-5001) as atest sample as shown in Table 14. A thirty member consumer panelevaluated two samples of lemon poppy seed muffins for several attributes(lemon, vanilla flavors, brown notes, sweet & bitter aftertaste).

TABLE 14 Effect of PCS-5001 (stevia extract) on baked goods SteviolSteviol Glycoside glycoside w/ (400 ppm) 120 ppm Ingredients Controlstevia extract DRY Ingredients Sucrose 12.3722 12.3682 All Purpose Flour17.6434 17.6434 Whole Wheat Flour 5.8763 5.8763 Poppy Seeds 1.06481.0648 Maltodextrin—10DE 2.1368 2.1368 Fibersol2 (ADM/Matsutani) 1.06481.0648 Modified Starch—Inscosity 656 1.0648 1.0648 LemonFlavor—Firmenich 0.8860 0.8860 Salt (Sodium Chloride) 0.7479 0.7479Baking Powder 1.0648 1.0648 Baking Soda 0.3205 0.3205 Steviol Glycoside0.0400 0.0400 Stevia extract (PCS-5001) 0.0120 Wet Ingredients Milk, 2%27.2444 27.2444 Soybean Oil 11.7525 11.7525 Whole Eggs 8.5473 8.5473Water 5.3420 5.3420 Yogurt, Plain Nonfat 1.6026 1.6026 Lemon Juice, 100%0.6410 0.6410 Vanilla Extract 0.5342 0.5342 100 100

FIG. 8 shows the modification of flavor and sweetness profiles caused bythe addition of Stevia extract (PCS-5001). The panel found that theaddition of Stevia extract provided an increase in brown note thancontrol sample without Stevia extract (at 90% confidence).

Example 8: Effect of Stevia Extract (PCS-5001) on Flavor Modification ofReduced Sodium Applications

A 30% salt reduced tortilla chip formulation was developed to determinethe flavor modification effect of Stevia extract (PCS-5001) in a saltreduced applications. To test the contribution of PCS-5001 in a saltreduced application, cheddar cheese flavor tortilla chips were coatedwith a control salt formulation, and a 30% salt reduced formulation withStevia extract (PCS-5001) as a test sample as shown in Table 15. Asixteen member consumer panel evaluated two samples of cheddar cheeseflavored tortilla chips for different attributes (sweet intensity,saltiness, cheese flavor, dairy notes, corn flavor, bitterness, andsweet & bitter aftertaste).

FIG. 9 shows the modification of flavor and salt perception caused bythe addition of Stevia extract (PCS-5001). The panel found the additionof Stevia extract in a 30% salt reduced formulation provided an increasein salt perception, parity to the full sodium control. In addition,Stevia extract provided an increase in sweet intensity and dairy notehigher than control sample without Stevia extract (at 95% confidence).

TABLE 15 Tortilla Chips with Cheddar Cheese 30% less sodium Control 30%Less Salt Corn chips 78 78.33 Cheese seasoning 10 10.04 Vegetable Oil 1111.05 Added Salt 1 0.57 PCS-5001 0.01 Total w (g) 100 100.00

Example 9: Effect of Stevia Extract (PCS-5001) on Flavor Modification ofDried Meat Applications

A beef jerky formulation was developed to determine the flavormodification effect of Stevia extract (PCS-5001) in a dried meatapplications. To test the contribution of PCS-5001 in a dried meatapplication, flank steak was marinated with a reduced sugar controlformulation, and a 30% sugar reduced formulation with steviol glycosidesand Stevia extract (PCS-5001) as a test sample as shown in Table 16. Atwenty member consumer panel evaluated two samples of beef jerky fordifferent attributes (sweet intensity, saltiness, black pepper, teriyakiflavor, fat-like intensity, beef flavor and sweet aftertaste).

FIG. 10 shows the modification of flavor and salt perception caused bythe addition of Stevia extract (PCS-5001). The panel found the additionof Stevia extract in a 30% sugar reduced formulation provided anincrease in salt perception.

TABLE 16 30% sugar reduced Beef Jerky Control (%) Stevia Extract FlankSteak 75.44 75.44 Balsamic vinegar 10.15 10.15 Salt 2.46 2.46 Pepper0.83 0.83 Sugar 6.88 6.88 Liquid smoke 0.86 0.86 Water Garlic powder0.44 0.44 Onion powder 0.44 0.44 Steviol Glycoside 0.018 0.018 PCS-5001(stevia extract) 0.0100 Worcestershire sauce 2.46 2.46 100 100

Example 10: Effect of Stevia Extract (PCS-5001) on Flavor Modificationof Reduced Sodium Applications in Brown Gravy

A 30% sodium reduced brown gravy formulation was developed to determinethe flavor modification effect of Stevia extract (PCS-5001) in a saltreduced applications. To test the contribution of PCS-5001 in a saltreduced application, a 30% sodium reduced brown gravy formulation, and a30% salt reduced formulation with Stevia extract (PCS-5001) as a testsample. A thirty member consumer panel evaluated two samples of browngravy for different attributes (sweet intensity, saltiness, blackpepper, beef flavor, and onion/savory notes, bitterness, and sweet &bitter aftertaste).

FIG. 11 shows the modification of flavor and salt perception caused bythe addition of Stevia extract (PCS-5001). The panel found the additionof Stevia extract in a 30% salt reduced formulation provided an increasein salt perception compared to 30% sodium reduced control. In addition,Stevia extract provided an increase in savory and black pepper notehigher than control sample without Stevia extract (at 95% confidence).There was also a decrease in bitter aftertaste.

Example 11: Effect of Stevia Extract on Flavor Modification of DairyProduct

To evaluate the contribution of PCS-1015 (MLD-1), a Stevia extract, to adairy product, two 50% reduced sugar chocolate milk samples wereprepared and tested by a consumer panel of 30 company employees. Theconsumer panel evaluated those two samples of chocolate milk for overallacceptance and attribute intensities (chocolate flavor, dairy notes,sweetness, bitterness and aftertaste) in two sessions. In session one,the two samples included: 1) a 50% sugar reduced control samplecontaining PureCircle Alpha (steviol glycoside sweetener) and 2) 50%sugar reduced test sample containing PureCircle Alpha and 70 ppmPCS-1015 (MLD-1). In session two, the two samples included: 1) a 50%sugar reduced control sample containing PureCircle Alpha (steviolglycoside sweetener) and 2) 50% sugar reduced test sample containingPureCircle Alpha and 80 ppm PCS-1015 (MLD-1). Tables 17 shows theformula of the control and test samples of 50% reduced sugar.

TABLE 17 50% sugar reduced Chocolate Milk with PCS-1015 50% Total 50%Total 50% Total Sugar Sugar Sugar Reduction Reduction Reduction withwith with PC PC PureCircle Alpha & Alpha & Dairy Formula Alpha PCS-1015PCS-1015 2% Reduced 96.5803 96.5743 96.5753 fat Milk Sugar 2.40 2.402.40 Cocoa Powder 0.80 0.80 0.80 10/12 Palsgaard 150 0.20 0.20 0.20ChoMilk PureCircle 0.0197 0.0197 0.0197 Alpha PCS-1015 0.0070 0.0080(MLD-1) Total 100 100 100

Table 18 shows the sensory results with the two test samples. Both testsamples showed the impact of the Stevia extract (PCS 1015) on theChocolate flavor notes and dairy note. At 80 ppm use level, thechocolate milk sample showed better sweetness profile and overall likingthan the control sample. FIG. 12 shows the comparison of the tasteprofile between the control and the test sample with 80 ppm Steviaextract PCS 1015.

TABLE 18 Summary of the overall acceptance and mean attribute intensityresults for each reduced sugar chocolate milk samples tested by 30 panelmembers. Summary of Mean-Scores, P-Values, and Significance Test ResultCode—chocolate milk with 70 ppm MLD-1 197 ppm 70 ppm of of Alpha MLD-1Only w/ PC Attribute (Control) Alpha P-Value Sig Sweet Intensity 8.858.89 0.8555 NS Chocolate Flavor 6.82 b 7.70 a 0.0482 *** Dairy Note 3.61b 4.19 a 0.1934 * Bitterness 0.84 0.83 0.9500 NS Bitter Aftertaste 0.740.70 0.6096 NS Sweet Aftertaste 3.02 3.15 0.7232 NS Overall Liking 7.127.42 0.5114 NS Summary of Mean-Scores, P-Values, and Significance TestResult Code—chocolate milk with 80 ppm MLD1 197 ppm 80 ppm of of AlphaMLD-1 only w/ PC Attribute (Control) Alpha P-Value Sig Sweet Intensity8.90 b 9.05 a 0.1557 * Chocolate Flavor 6.89 b 7.53 a 0.0048 *** DairyNote 4.12 b 4.44 a 0.1470 * Bitterness 0.49 0.35 0.2473 NS BitterAftertaste 0.71 a 0.55 b 0.1824 * Sweet Aftertaste 2.66 2.82 0.5177 NSOverall Liking 6.49 b 6.89 a 0.1908 * * = 80% CI, ** = 90% CI, *** = 95%CI

Example 12: Effect of Stevia Extract on Desserts (Vanilla Custard)

To test the contribution of the Stevia extract, PCS-1015 in gelatin andpuddings, two 30% calorie reduced vanilla custard samples weretested: 1) sweetened with PureCircle Alpha, a PureCircle Steviasweetener, 2) sweetened with PureCircle Alpha and PCS-1015 (MLD-1).Table 19 shows the formulation of the control and test samples. A panelof 30 trained panelists with extensive experience in profiling sensoryattributes tasted both samples.

To prepare the sample, blend the PureCircle Alpha and the testingredient (PCS-1015) with the dry ingredients. Add the dry ingredientsto the milk using good agitation. Heat on low until all ingredients aredissolved. Heat up to 95° C. for 10 minutes to cook up the starches. Addflavors, stir it, cool, stir it before place it in the refrigerator.Serve at chilled in 1 oz cups.

TABLE 19 Reduced sugar dessert (Vanilla Custard) with PCS-1015 Controlwith Test with PureCircle PureCircle Alpha Alpha w/ stevia extract Milk( 1% fat) 94.27 94.27 Sucrose 4.00 4.00 Starch Perma Flo 1.25 1.25 Tate& Lyle TIC Carrageenan 0.09 0.09 Salt 0.06 0.06 ROHA Beta Carotene 0.050.05 French Vanilla Flavor 0.15 0.15 UV 420-066-7 Steviol Glycoside0.0166 0.0166 Stevia Extract — 0.0080 Total 100 100

The trained panel found that the test sample had stronger sweetintensity, vanilla, dairy flavor notes and overall liking at 80%confidence. The sample containing Stevia extract also had significantlyhigher egg note at 95% confidence. FIG. 13 shows the pictorial renditionof the sensory difference between the control and test dessert samples

TABLE 20 Summary of the overall acceptance and mean attribute intensityresults for reduced sugar dessert (Vanilla Custard) with PCS-1015 166ppm of 70 ppm Alpha of Only MLD-1 Attribute (Control) with Alpha P-ValueSig Sweet Intensity 7.01 a 7.13 b 0.1095 * Vanilla Flavor 3.22 a 3.5 b0.1299 * Egg Note 1.22 a 1.56 b 0.0497 *** Dairy / Creaminess 3.04 a3.22 b 0.1164 * Bitterness 0.43 0.5 0.3001 NS Bitter Aftertaste 0.360.38 0.7692 NS Sweet Aftertaste 2.23 2.24 0.8794 NS Overall Liking 6.49a 6.87 b 0.1149 *

Example 13: Effect of Stevia Extract on Flavor Modification ofChocolate-Flavored Beverage with Cocoa Powder Reduction

A chocolate flavored dairy beverage was developed to determine theflavor modification effect of Stevia extract flavor with modifyingproperties (FMP) in a dairy beverage. The two samples included: 1)control sample with full amounts of sugar and cocoa powder, and 2) testsample with 15% reduced sugar and 20% reduced cocoa, containing 60 ppmof Stevia extract FMP, as shown in Table 21.

TABLE 21 Reduced Sugar and Cocoa Chocolate-Flavored Beverage (ChocolateMilk) with Stevia Extract Test with Control with Reduced Full AmountSugar and of Sugar Cocoa with and Cocoa Stevia Extract Milk, 2% milkfat85.200 86.554 Sugar 8.00 6.80 Hot Water, 190° F. 6 6 Natural 10/12 0.800.64 Cocoa Powder Stevia Extract — 0.006 FMP (PSB-5005) Total 100 100

A 15 member trained panel evaluated samples of chocolate milk foroverall acceptance and attribute intensities (sweet intensity,bitterness, cocoa flavor, dairy note, sweet aftertaste and bitteraftertaste). The parameters for the sensory evaluation are shown inTable 22.

Table 23 shows the sensory results for the control and test products.The test product with 20% reduced cocoa powder and Stevia extract FMPshows no significant difference in cocoa flavor from the control. Thetest sample with 60 ppm Stevia extract FMP was higher in sweet intensity(90% confidence) and sweet aftertaste (directional). FIG. 14 illustratesthis comparison.

TABLE 22 Sensory evaluation of chocolate flavored beverage Nature ofParticipants: Trained panel Number of Sessions  1 Number ofParticipants: 15 Test Design: Balanced, randomized within set. BlindSensory Test Method: Intensity and acceptance ratings EnvironmentalCondition Standard booth lighting Attributes and Scales: OverallAcceptance on a 10-pt hedonic scale where 10 = Extremely Like and 0 =Extremely Dislike Overall Liking, sweet intensity, bitterness, cocoaflavor, dairy note, sweet aftertaste and bitter aftertaste. 10-ptcontinuous intensity scale where 0 = Imperceptible and 10 = ExtremelyPronounced Statistical Analysis: ANOVA (by Block) with Post Hoc Duncan'sTest Sample Size ~1.5 oz. in a clear capped plastic cup ServingTemperature Refrigerated temperature (~45° F.) Serving/Panelists Samplesserved simultaneously. Instruction: Panelists instructed to evaluateeach sample.

TABLE 23 Sensory Results Test with Control Reduced with Full Sugar andAmount of Cocoa with Sugar and Stevia Attribute Cocoa Extract FMP Sweetintensity 7.05 7.26 Bitterness 0.65 0.55 Cocoa flavor 4.4 4.28 Dairynote 4.91 5.13 Sweet aftertaste 1.24 1.43 Bitter aftertaste 0.21 0.16Overall liking 7.43 7.12

As seen in Table 23, the test product with reduced sugar and cocoa, andcontaining Stevia extract FMP, in this case PSB-5005, had statisticallysimilar overall liking and mean cocoa flavor intensity results ascompared to a full-cocoa formulation. The test product containing Steviaextract FMP had lower bitterness attribute and bitter aftertaste ratingscompared to the control product made without Stevia extract. The dairynote was rated higher in the test product compared to the controlproduct. From these results it can be seen that a reduction in cocoa andsugar content in a dairy beverage can be suitably accomplished using aStevia extract FMP, such as PSB-5005, and unexpectedly with a decreasein bitterness which is typically associated with Stevia ingredients.

Example 14: Effect of Stevia Extract FMP on Flavor Modification ofVanilla-Flavored Dairy Product

A 50% sugar-reduced vanilla yogurt was developed to determine the flavormodification effect of Stevia extract flavor with modifying properties(FMP) in a reduced-sugar vanilla-flavored dairy product. The two samplesas shown in Table 24 included: 1) control sample with 180 ppm steviolglycoside sweetener, and 2) test sample with 180 ppm steviol glycosidesweetener and 100 ppm of Stevia extract FMP.

TABLE 24 Sugar Reduced Vanilla-Flavored Dairy Product (Vanilla Yogurt)Test with Control Reduced 180 ppm Sugar and steviol Cocoa with glycosideStevia sweetener Extract FMP Plain nonfat yogurt 96.132 96.122 Sugar3.750 3.750 Vanilla Flavor 0.100 0.100 Steviol Glycoside 0.018 0.018Stevia Extract — 0.010 (PCS 5001) Total 100.000 100.000

A 30 member panel evaluated samples of vanilla yogurt for overallacceptance and attribute intensities (sweet intensity, bitterness,vanilla flavor, dairy, astringency, sweet aftertaste and bitteraftertaste). Table 25 lists the sensory evaluation parameters.

TABLE 25 Sensory evaluation of vanilla flavored dairy product Nature ofParticipants: Trained sensory panel Number of Sessions  1 Number ofParticipants: 30 Test Design: Balanced, randomized within set. BlindSensory Test Method: Intensity and acceptance ratings EnvironmentalCondition Standard booth lighting Attributes and Scales: OverallAcceptance on a 10-pt hedonic scale where 10 = Extremely Like and 0 =Extremely Dislike Overall Liking, sweet intensity, bitterness, vanillaflavor, dairy, astringency, sweet aftertaste and bitter aftertaste.10-pt continuous intensity scale where 0 = Imperceptible and 10 =Extremely Pronounced Statistical Analysis: ANOVA (by Block) with PostHoc Duncan's Test Sample Size ~1.5 oz. in a clear capped plastic cupServing Temperature Refrigerated temperature (~45° F.) Serving/PanelistsSamples served simultaneously. Instruction: Panelists instructed toevaluate each sample.

Table 26 shows the sensory results for the control and test products. At95% confidence, the test sample containing Stevia extract FMP wassignificantly higher for sweet intensity and vanilla flavor andsignificantly and unexpectedly lower in bitterness, astringency andsweet aftertaste. At 90% confidence, the test sample was higher in dairyand had higher overall liking. FIG. 15 illustrates this comparison.

TABLE 26 Summary of the overall acceptance and mean attribute intensityresults for vanilla-flavored dairy product (vanilla yogurt) with steviaextract Test with 180 ppm steviol Control glycoside 180 ppm sweetenerand steviol 100 ppm glycoside stevia extract Attribute sweetener FMPp-value Sig Sweet intensity 6.88 7.05 0.0281 *** Bitterness 2.53 1.740.0374 *** Vanilla flavor 4.95 5.87 0.0452 *** Dairy 4.78 5.65 0.0507 **Astringency 2.36 1.63 0.0407 *** Sweet aftertaste 1.73 0.95 0.0144 ***Bitter aftertaste 1.55 0.94 0.0682 ** Overall liking 5.26 6.37 0.0629 **

Example 15: Usage Levels of Stevia Extract FMP

Useful or maximum usage levels of Stevia extract FMPs were evaluated. Tobe useful as a flavor with modifying properties, the level of use of theStevia extract must be below a certain sweetness detection threshold ina particular food or beverage product. To determine this threshold, asensory evaluation is conducted with a full sugar product as thecontrol, and a test product containing different levels of the Steviaextract FMP. Sensory panel members are then asked to identify whichproduct is sweeter.

Using the Flavor and Extract Manufacturers' Association (FEMA) guidancedocument called “Guidance for the Sensory Testing of Flavorings withModifying Properties within the FEMA GRAS™ Program, 2013”, therecognition threshold was determined using a 2-alternatve forced choice(2-AFC) methodology, as described in Table 27.

TABLE 27 Sensory evaluation to determine usage levels Nature ofParticipants: Company employees Number of Sessions  1 Number ofParticipants: 30 Test Design: 2- AFC, Balanced, randomized within pair.Blind Sensory Test Method: Intensity ratings Environmental ConditionStandard booth lighting Attributes and Scales: Which sample is sweeter?Statistical Analysis: Paired comparison Test Sample Size ~1.5 oz. in aclear capped plastic cup Serving Temperature Refrigerated temperature(~42° F.) or room temperature, depending on sample requirementsServing/Panelists Samples served simultaneously. Instruction: Panelistsinstructed to read ingredient statement, evaluate each sample.

Usage levels for Stevia extract FMP are determined by those levels atwhich the Stevia extract FMP provides a sweetness perception that issignificantly less than the full sugar control. For products other thanbaked goods and breakfast cereals, the sugar level in the controlproduct was 1.5%. In baked goods, the sugar level in the control productwas 4%, and in breakfast cereals, the sugar level in the control productwas 3%. The test products contained no added sugar and contain variouslevels of Stevia extract FMP.

Table 28 shows usage levels of Stevia extract FMP in various food andbeverage applications as determined using the FEMA sensory testingguidance.

TABLE 28 Usage levels of stevia extract FMP Category Usage Level (ppm)Baked Goods 500 Beverages, Non-Alcoholic 110 Beverages, Alcoholic 130Breakfast Cereals 600 Chewing Gum 100 Condiments and Relishes 100Confections and Frostings 100 Fats and Oils 180 Frozen Dairy 100 FruitIces 100 Gelatins and Puddings 100 Gravies 100 Hard Candy 100 ImitationDairy 165 Instant Coffee and Tea 200 Jams and Jellies 100 Milk Products165 Nut Products 230 Processed Fruits 100 Processed Vegetables 100Seasonings and Flavors 230 Snack Foods 230 Soft Candy 100 Soups 100Sugar Substitutes 100 Sweet Sauces 100

It was unexpectedly discovered that Stevia extract FMPs can be used atvarious levels to favorably impact the taste and flavor profile of afood or beverage product while having little or no detectable sweetnessperception in that product. These usage levels serve as examples of use,and other usage levels of the Stevia extract FMP in various consumableproducts are contemplated by this invention.

Although various embodiments of the present invention have beendisclosed here for purposes of illustration, it should be understoodthat a variety of changes, modifications and substitutions may beincorporated without departing from either the spirit or the scope ofthe invention.

1. A Stevia extract flavor with modifying properties (FMP) comprising:(a) rebaudioside A at a level of up to about 30% by weight; (b)stevioside at a level of up to about 12% by weight; and (c) minorsteviol glycosides and plant molecules at a level of up to 55% byweight, wherein the Stevia extract FMP modifies the flavor and sweetnessprofile of a consumable product without imparting significant sweetnessto the consumable product.
 2. The Stevia extract FMP of claim 1, furthercomprising up to about 25% rebaudioside C by weight and up to about 20%rebaudioside D by weight.
 3. The Stevia extract FMP of claim 1, furthercomprising up to about 6% rebaudioside E by weight, up to about 8%rebaudioside N by weight, and up to about 8% rebaudioside O by weight.4. The Stevia extract FMP of claim 1, wherein the minor steviolglycosides comprise one or more selected from the group consisting of:rebaudioside G; rebaudioside H; rebaudioside I, rebaudioside J;rebaudioside K; and rebaudioside L.
 5. The Stevia extract FMP of claim1, wherein the plant glycosides comprise one or more selected from thegroup consisting of: glucosyl-4′-O-apigenin; glucosyl-7-O-luteolin;rhamnosyl-3-O-kaempferol; glucosyl-3-O-quercitin; andarabinosyl-3-O-quercetin. 6.-11. (canceled)